Nodavirus-Based Vaccines for West Nile Virus (pilot)

基于诺达病毒的西尼罗河病毒疫苗（试点）

• 批准号: 7283450
• 负责人: KYLE L JOHNSON
• 金额: $ 7.32万
• 依托单位: UNIVERSITY OF TEXAS EL PASO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-06-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7283450
• 关键词: Animals; Antibodies; Arthropods; Bioterrorism; Bos taurus structural-GP protein; Categories; Cells; Development; Disease; Goals; Health Priorities; Hepatitis B Vaccines; Human; Human Virus; Immune response; Insecta; Lead; Mammalian Cell; Mammals; Mediating; Messenger RNA; Mission; Mus; National Institute of Allergy and Infectious Disease; PTPN11 gene; Pathogenicity; Plants; Proteins; Public Health; RNA; RNA replication; Rate; Replicon; Research; Research Project Grants; Saccharomyces cerevisiae; Source; System; Testing; Transfection; United States; United States National Institutes of Health; Vaccines; Virus; West Nile virus; Yeasts; aerosolized; base; biodefense; cell mediated immune response; concept; immunogenicity; mortality; novel; pathogen; prevent; transmission process; viral RNA

West Nile virus (WNV) is a serious human pathogen with a mortality rate of 0.1%. The National Institute for Allergy and Infectious Disease (NIAID) Biodefense Research Agenda classifies WNV as a Category B Priority Pathogen on the basis of its transmission by an arthropod host, its infectivity on aerosolized exposure, and its potential threat to public health. Of particular concern is the possibility that West Nile virus might be used as an agent of bioterrorism. There is currently no vaccine for WNV available for use in humans. Thus, development of a WNV vaccine is a national biodefense and public health priority that is consistent with the overall goals of NIH and MBRS. The goals of this Pilot Research Project are to develop a candidate vaccine for West Nile virus and to test its immunogenicity in mice. This proposal incorporates two novel concepts: the use of Nodamura virus (NoV) RNA replicons to amplify WNV mRNAs in the yeast Saccharomyces cerevisiae and inoculation of animals with purified total yeast RNA containing amplified NoV-WNV RNAs. The advantages of the nodavirus expression system as a source of vaccine material include exponential amplification of the chimeric mRNA; its ability to initiate RNA replication in a wide variety of host cells, including those from mammals, insects, plants, and yeast, when the RNA is introduced by transfection; and the lack of pathogenicity of NoV for humans. Yeast cells provide a safe and inexpensive source of vaccine material, and are currently used to produce Hepatitis B vaccines. The specific aims of this proposal are: 1. Construct NoV RNA2-based replicons that contain WNV structural (glycoprotein E, gpE) or nonstructural (NS1) proteins and determine the extent to which RNA replication amplifies NoV2-gpE and NoV-NS1 mRNA and protein levels in yeast and mammalian cells 2. Evaluate the WNV structural (gpE) and nonstructural (NS1) proteins as potential vaccine candidates by defining the humoral and cell-mediated immune responses to these proteins elicited in inoculated mice. Relevance to public health: West Nile virus is a severe human pathogen that poses a threat to the public health and biosafety of the United States. Our goal is to develop safe, effective candidate vaccines that will prevent the spread of West Nile virus-associated diseases.

西尼罗河病毒（WNV）是一种严重的人类病原体，死亡率为0.1%。国家研究所 过敏和传染病 (NIAID) 生物防御研究议程将西尼罗河病毒列为 B 类 优先病原体基于其通过节肢动物宿主的传播、其对雾化的传染性 暴露及其对公共健康的潜在威胁。特别值得关注的是西尼罗河病毒的可能性 可能被用作生物恐怖主义的代理人。目前尚无西尼罗河病毒疫苗可供使用 人类。因此，开发西尼罗河病毒疫苗是国家生物防御和公共卫生的优先事项 与 NIH 和 MBRS 的总体目标一致。该试点研究项目的目标是 开发西尼罗河病毒候选疫苗并在小鼠中测试其免疫原性。这个提议 融合了两个新颖的概念：使用 Nodamura 病毒 (NoV) RNA 复制子来扩增 WNV mRNA 酿酒酵母和用含有纯化总酵母RNA的动物接种 扩增的 NoV-WNV RNA。诺达病毒表达系统作为疫苗来源的优势 材料包括嵌合mRNA的指数扩增；其启动 RNA 复制的能力 当 RNA 存在于多种宿主细胞中时，包括来自哺乳动物、昆虫、植物和酵母的宿主细胞 通过转染引入； NoV 对人类缺乏致病性。酵母细胞提供安全且 疫苗材料的廉价来源，目前用于生产乙型肝炎疫苗。这 该提案的具体目标是： 1. 构建基于 NoV RNA2 的复制子，其中包含 WNV 结构（糖蛋白 E、gpE）或非结构 (NS1) 蛋白并确定 RNA 复制放大 NoV2-gpE 和 NoV-NS1 的程度 酵母和哺乳动物细胞中的 mRNA 和蛋白质水平 2. 通过以下方法评估 WNV 结构 (gpE) 和非结构 (NS1) 蛋白作为潜在候选疫苗的可能性 定义了在接种小鼠中引发的对这些蛋白质的体液和细胞介导的免疫反应。 与公共卫生的相关性：西尼罗河病毒是一种严重的人类病原体，对公众构成威胁 美国的健康和生物安全。我们的目标是开发安全、有效的候选疫苗 防止西尼罗河病毒相关疾病的传播。